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. 2021 Oct 20;12:692431. doi: 10.3389/fphar.2021.692431

FIGURE 4.

FIGURE 4

Effects of an autophagy inhibitor on autophagy and apoptosis in HASMCs. After pretreatment with wortmannin (WM) for 1 h, HASMCs were incubated with AGEs (100 µg/ml) for another 24 h (A) WM suppressed AGEs-induced autophagy marker expression in HASMCs. Western blot analysis of autophagy proteins (LC3, beclin1, p62, and p-mTOR) in HASMCs treated with WM and AGEs for 24 h (B) WM abolished AGEs-induced autophagic puncta formation in HASMCs. Immunofluorescence analysis of LC3-II puncta formation (red color represents LC3-II, blue represents DAPI; scale bar = 25 μm). (C) TEM evidence of autophagic vacuoles (red arrows), bars: 1 μm. (D) Western bDlot analysis of apoptosis-related proteins (Bak, Bax, caspase-3, and caspase-9) in WM/AGEs -treated HASMCs. Data are presented as fold change compared with the control group. Each point represents the mean ± SD (n = 3/group); * p < 0.05, ** p < 0.01, *** p < 0.001 compared with the WM/AGEs-treated group; # p < 0.05, ## p < 0.01, ### p < 0.001.